PDF(Pubmed)
Abstract：
In the human body, proteins secreted into peripheral blood vessels are known as the secretome, and they represent the physiological or pathological status of cells. The unique response of cells to toxin exposure can be confirmed via secretome analysis, which can be used to discover toxic mechanisms or exposure markers. Alpha-amanitin (α-AMA) is the most widely studied amatoxin and inhibits transcription and protein synthesis by directly interacting with RNA polymerase II. However, secretory proteins released during hepatic failure caused by α-AMA have not been fully characterized. In this study, we analyzed the secretome of α-AMA-treated Huh-7 cells and mice using a comparative proteomics technique. Overall, 1440 and 208 proteins were quantified in cell media and mouse serum, respectively. Based on the bioinformatics results for the commonly downregulated proteins in cell media and mouse serum, we identified complement component 3 (C3) as a marker for α-AMA-induced hepatotoxicity. Through western blot in cell secretome and C3 ELISA assays in mouse serum, we validated α-AMA-induced downregulation of C3. In conclusion, using comparative proteomics and molecular biology techniques, we found that α-AMA-induced hepatotoxicity reduced C3 levels in the secretome. We expect that this study will aid in identifying new toxic mechanisms, therapeutic targets, and exposure markers of α-AMA-induced hepatotoxicity.
UNASSIGNED: The online version contains supplementary material available at 10.1007/s43188-022-00163-z.
UNASSIGNED: The online version contains supplementary material available at 10.1007/s43188-022-00163-z.
摘要：
在人体中,分泌到外周血管的蛋白质被称为分泌组，它们代表细胞的生理或病理状态。细胞对毒素暴露的独特反应可以通过分泌组分析来证实，可用于发现毒性机制或暴露标志物。α-amanitin（α-AMA）是研究最广泛的amatoxin，通过与RNA聚合酶II直接相互作用来抑制转录和蛋白质合成。然而,在由α-AMA引起的肝衰竭期间释放的分泌蛋白尚未被完全表征。在这项研究中,我们使用比较蛋白质组学技术分析了α-AMA处理的Huh-7细胞和小鼠的分泌组。总的来说,1440和208蛋白质在细胞培养基和小鼠血清中定量，分别。基于细胞培养基和小鼠血清中常见下调蛋白的生物信息学结果，我们确定补体成分3（C3）是α-AMA诱导的肝毒性的标志物。通过细胞分泌组中的蛋白质印迹和小鼠血清中的C3ELISA测定，我们验证了α-AMA诱导的C3下调。总之,使用比较蛋白质组学和分子生物学技术，我们发现α-AMA诱导的肝毒性降低了分泌组中的C3水平。我们希望这项研究将有助于确定新的毒性机制，治疗目标，和α-AMA诱导的肝毒性的暴露标志物。
■在线版本包含补充材料，可在10.1007/s43188-022-00163-z获得。
■在线版本包含补充材料，可在10.1007/s43188-022-00163-z获得。
